LOCUS       sequence-315434-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a ST6GAL1 mutant with a GFP tag in its C-terminus in 
            mammalian cells. The mutated stem region of Tac is inserted within 
            ST..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  AUTHORS   Sun X, Tie HC, Chen B, Lu L
  TITLE     Glycans function as a Golgi export signal to promote the 
            constitutive exocytic trafficking.
  JOURNAL   J Biol Chem. 2020 Aug 21. pii: RA120.014476. doi: 
            10.1074/jbc.RA120.014476.
  PUBMED    32826314
REFERENCE   2  (bases 1 to 6005)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2020 Aug 21. pii: RA120.014476. doi: 
            10.1074/jbc.RA120.014476."
COMMENT     Sequence Label: ST-Tstem(5A)-GFP
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1945..1954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1951..1953,1954..1956,1957..2670)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1996..2017)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2793..2914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2830..2849)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2884..2903
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2921..3376)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3079)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3270..3289
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3403..3507
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3509..3866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3509..3700
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3532..3552)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3717..3852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3901..4695
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3901..4695
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3955..3974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4565..4584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4883..4902)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4927..4974
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5303..5891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5792..5811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgatt cacaccaacc tgaagaaaaa gttcagctgc
      661 tgcgtcctgg tctttcttct gtttgcagtc atctgtgtgt ggaaggaaaa gaagaaaggg
      721 agttactatg attcctttaa attgcaaacc aaggaattcc aggtgttaaa gagtctgggg
      781 aaattggcca tggggtctga ttccccgcgg ctcgtcacaa cagcagattt tcaaatacag
      841 gcagaaatgg ctgcagccat ggagacgtcc atatttgcag cagagtacca gggatcccag
      901 tctgtatcct caagcagcac ccaggacccc cacaggggcc gccagaccct cggcagtctc
      961 agaggcctag ccaaggccaa accagaggcc tccttccagg tgtggaacaa ggacagctct
     1021 tccaaaaacc ttatccctag gctgcaaaag atctggaaga attacctaag catgaacaag
     1081 tacaaagtgt cctacaaggg gccaggacca ggcatcaagt tcagtgcaga ggccctgcgc
     1141 tgccacctcc gggaccatgt gaatgtatcc atggtagagg tcacagattt tcccttcaat
     1201 acctctgaat gggagggtta tctgcccaag gagagcatta ggaccaaggc tgggccttgg
     1261 ggcaggtgtg ctgttgtgtc gtcagcggga tctctgaagt cctcccaact aggcagagaa
     1321 atcgatgatc atgacgcagt cctgaggttt aatggggcac ccacagccaa cttccaacaa
     1381 gatgtgggca caaaaactac cattcgcctg atgaactctc agttggttac cacagagaag
     1441 cgcttcctca aagacagttt gtacaatgaa ggaatcctaa ttgtatggga cccatctgta
     1501 taccactcag atatcccaaa gtggtaccag aatccggatt ataatttctt taacaactac
     1561 aagacttatc gtaagctgca ccccaatcag cccttttaca tcctcaagcc ccagatgcct
     1621 tgggagctat gggacattct tcaagaaatc tccccagaag agattcagcc aaacccccca
     1681 tcctctggga tgcttggtat catcatcatg atgacgctgt gtgaccaggt ggatatttat
     1741 gagttcctcc catccaagcg caagactgac gtgtgctact actaccagaa gttcttcgat
     1801 agtgcctgca cgatgggtgc ctaccacccg ctgctctatg agaagaattt ggtgaagcat
     1861 ctcaaccagg gcacagatga ggacatctac ctgcttggaa aagccacact gcctggcttc
     1921 cggaccattc actgcgcacc ggtcgccacc atggtgtcca agggcgagga gctgttcacc
     1981 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttctccgtg
     2041 cggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catcagcacc
     2101 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     2161 agcttctccc gctaccccga ccacatgaag cgccacgact tcttcaagag cgccatgccc
     2221 gaaggctacg tccaggagcg caccatctcc ttcaaggacg acggcaccta caagacccgc
     2281 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2341 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa ctcccacaac
     2401 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     2461 aacgtggagg acggctccgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2521 gacggccccg tgctgctgcc cgacaaccac tacctgtcca cccagtccaa gctgtccaaa
     2581 gaccccaacg agaagcgcga tcacatggtc cttctggaat tcgtgaccgc cgccgggatc
     2641 actcacggca tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc
     2701 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2761 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2881 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2941 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3001 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3061 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3121 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3181 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3241 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3301 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3361 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3421 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3481 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3541 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3841 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4741 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4801 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4861 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4921 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4981 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5041 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5101 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6001 tgcat
//
